Tatsuaki Tsuruyama, Tonau Nakai, Takuya Hiratsuka, Guang Jin, Takuro Nakamura and Kenichi Yoshikawa
Scientific Tracks Abstracts: JAA
Although there have been a few reports that the HIV-1 genome can be selectively integrated into the genomic DNA of cultured host cell, the biochemistry of integration selectivity has not been fully understood. We modifi ed the in vitro integration reaction protocol and developed a reaction system with higher effi ciency. We used a substrate repeat, 5ï¿½- (GTCCCTTCCCAGT) n(ACTGGGAAGGGAC)n-3ï¿½, and a modifi ed sequence DNA ligated into a circular plasmid. CAGT and ACTG in the repeat units originated from the HIV-1 proviral genome ends. Following the incubation of the HIV-1 genome end cDNA and recombinant integrase for the formation of the preintegration (PI) complex, substrate DNA was reacted with this complex. As a result, it was confi rmed that the integration selectively occurred. CAGT motif and DNA secondary structure in the target sequence were cardinal factors for the selective In conclusion, there is a considerable selectivity in HIV-integration into the specifi ed sequence; however, similar DNA sequences can interfere with the integration process, and it is therefore diffi cult for in vivo integration to occur selectively in the actual host genome DNA.
Researcher: Department of Pathology, Kyoto University, Faculty of Medicine & Kyoto University Hospital (1995- 1998)Lecturer: Department of Pathology, Kyoto University, Faculty of Medicine & Kyoto University Hospital (1998-2005)Associate Professor: Department of Pathology, Kyoto University, faculty of Medicine & Kyoto University Hospital (2005-).